In situ hybridization (Both Radioactive and Nonradioactive) and spatiotemporal gene expression analysis

Olga Simmons, Esther M. Bolanis, Jian Wang, Simon J. Conway

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

Abstract

Section in situ hybridization using either radioactive or nonradioactive labeled cDNA probes is an invaluable technique that enables the investigator to detect and localize mRNA expression within tissue sections and cells. Here, we describe the labeling of <sup>35</sup>S-UTP radioactive and nonradioactive digoxigenin probes, preparation of tissue sections, hybridization, and washing of non-hybridized probes, followed by the detection of radioactive signals via dipping in nuclear emulsion and the immunohistochemical and subsequent colorimetric detection of nonradioactive signals.

Original languageEnglish (US)
Title of host publicationMouse Genetics
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages225-244
Number of pages20
ISBN (Print)9781493912148
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1194
ISSN (Print)1064-3745

Keywords

  • cDNA probes
  • Digoxigenin
  • Immunohistochemistry
  • mRNA localization
  • Radioactive and nonradioactive in situ hybridization
  • Tissue sections

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Simmons, O., Bolanis, E. M., Wang, J., & Conway, S. J. (2014). In situ hybridization (Both Radioactive and Nonradioactive) and spatiotemporal gene expression analysis. In Mouse Genetics: Methods and Protocols (pp. 225-244). (Methods in Molecular Biology; Vol. 1194). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-1215-5_12